{"created":"2023-06-19T10:21:39.758961+00:00","id":636,"links":{},"metadata":{"_buckets":{"deposit":"ce01baa5-eba6-4eb6-9d09-676e01fdc084"},"_deposit":{"created_by":12,"id":"636","owners":[12],"pid":{"revision_id":0,"type":"depid","value":"636"},"status":"published"},"_oai":{"id":"oai:saitama-med.repo.nii.ac.jp:00000636","sets":["6:10:90"]},"author_link":["2185","2184"],"item_10006_date_granted_11":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"2018-03-23"}]},"item_10006_degree_grantor_9":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_name":"埼玉医科大学"}]}]},"item_10006_degree_name_8":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"博士(医学)"}]},"item_10006_description_10":{"attribute_name":"学位授与年度","attribute_value_mlt":[{"subitem_description":"平成29年度","subitem_description_type":"Other"}]},"item_10006_description_7":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Rapid and easy detection of a single nucleotide point mutation of bacterial genes, which is directly linked to drug susceptibility, is essential for the proper use of antimicrobial agents. Here, we established a detection method using a peptide nucleic acid mediated loop-mediated amplification (LAMP) assay for macrolide (ML)-susceptible Mycoplasma pneumoniae. This assay specifically detected the absence of missense mutations encoding the central loop of domain V in the gene encoding 23S rRNA, which can reduce the affinity for MLs and subsequently generate ML-resistant strains of M. pneumoniae. Reactions were performed at 62°C for 60min and targeted gene amplifications were detected by real-time turbidity with a turbidimeter and naked-eye inspection of a color change. The assay had an equivalent detection limit of 100.0fg of DNA with the turbidimeter and showed specificity against 54 types of pathogens, whereas amplification was completely blocked, even at 100.0pg of DNA per reaction, in the presence of point mutations at 2063A and 2064A. The expected LAMP products were confirmed through identical melting curves in real-time LAMP procedures. This method would be a simple and rapid protocol for single nucleotide polymorphism genotyping as point-of-care testing technology without amplification of the sequences carrying the point mutations 2063A and 2064A in ML-resistant M. pneumoniae strains.","subitem_description_type":"Abstract"}]},"item_10006_dissertation_number_12":{"attribute_name":"学位授与番号","attribute_value_mlt":[{"subitem_dissertationnumber":"32409甲第1384号"}]},"item_10006_relation_14":{"attribute_name":"掲載誌名","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"Journal of Microbiological Methods"}]}]},"item_10006_relation_15":{"attribute_name":"掲載誌情報","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"出版社サイト"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://www.sciencedirect.com/science/article/pii/S0167701217302178?via%3Dihub","subitem_relation_type_select":"URI"}}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"酒井, 純"},{"creatorName":"サカイ, ジュン","creatorNameLang":"ja-Kana"}],"nameIdentifiers":[{"nameIdentifier":"2184","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Sakai, Jun","creatorNameLang":"en"}],"nameIdentifiers":[{"nameIdentifier":"2185","nameIdentifierScheme":"WEKO"}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2018-05-30"}],"displaytype":"detail","filename":"1384-1.pdf","filesize":[{"value":"152.3 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"論文内容の要旨","url":"https://saitama-med.repo.nii.ac.jp/record/636/files/1384-1.pdf"},"version_id":"42c46886-96bb-4a69-b078-a7d17d6147ba"},{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2018-05-30"}],"displaytype":"detail","filename":"1384-2.pdf","filesize":[{"value":"81.1 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"論文審査の結果の要旨","url":"https://saitama-med.repo.nii.ac.jp/record/636/files/1384-2.pdf"},"version_id":"c0f54cf3-9b4d-48de-93b6-e44196a40fe9"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"Colorimetry","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"DNA, Bacterial","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Genes, Bacterial","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Genes, rRNA","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Genotype","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Humans","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Limit of Detection","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Mycoplasma pneumoniae","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Nasopharynx","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Nucleic Acid Amplification Techniques","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Peptide Nucleic Acids","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Pneumonia, Mycoplasma","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Point Mutation","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Point-of-Care Testing","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Polymerase Chain Reaction","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"RNA, Ribosomal, 23S","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Sensitivity and Specificity","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Temperature","subitem_subject_language":"en","subitem_subject_scheme":"Other"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"thesis","resourceuri":"http://purl.org/coar/resource_type/c_46ec"}]},"item_title":"ペプチド核酸peptide nucleic acid (PNA)とloop-mediated isothermal amplification assay（LAMP）法を組み合わせた、Mycoplasma pneumoniaeの23S rRNA変異の革新的検出法","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"ペプチド核酸peptide nucleic acid (PNA)とloop-mediated isothermal amplification assay（LAMP）法を組み合わせた、Mycoplasma pneumoniaeの23S rRNA変異の革新的検出法"},{"subitem_title":"A novel detection procedure for mutations in the 23S rRNA gene of Mycoplasma pneumoniae with peptide nucleic acid-mediated loop-mediated isothermal amplification assay.","subitem_title_language":"en"}]},"item_type_id":"10006","owner":"12","path":["90"],"pubdate":{"attribute_name":"公開日","attribute_value":"2018-05-30"},"publish_date":"2018-05-30","publish_status":"0","recid":"636","relation_version_is_last":true,"title":["ペプチド核酸peptide nucleic acid (PNA)とloop-mediated isothermal amplification assay（LAMP）法を組み合わせた、Mycoplasma pneumoniaeの23S rRNA変異の革新的検出法"],"weko_creator_id":"12","weko_shared_id":-1},"updated":"2023-06-19T10:33:57.306198+00:00"}